蟾蜍灵对HL—60细胞的生长抑制及凋亡诱导作用

目的 研究中药蟾酥有效成分蟾蜍灵（ｂｕｆａｌｉｎ）对人白血病细胞的作用及其机制。方法 应用ＭＴＴ比色法观察蟾蜍灵对细胞的抑制作用。荧光显微境和透射电镜观察细胞结果色的改变，ＤＮＡ交电泳１、原位缺口标记法和流式细胞术等分析细胞凋亡。结果 ①蟾蜍灵明显抑制ＨＬ－６０细胞的生长，半数抑制浓度（ＩＣ５０）约为０．０２５μｍｏｌ／Ｌ；②典型的细胞形态学改变、ＤＮＡ片段化和亚Ｇ１峰的检出等证实蟾蜍灵能诱导白血

The mechanisms of inhibitory effect of bufalin on human leukemia cells

OBJECTIVE: To study the mechanism of bufalin on human leukemia cell inhibition. METHODS: HL-60 cells were treated with bufalin at different concentrations. The growth inhibition was analysed by MTT assay, cell apoptosis by light microscopy, transmission electron microscopy, flow cytometry, TUNEL labeling method and agarose gel electrophoresis. RESULTS: (1) Treatment of HL-60 cells with bufalin remarkably inhibited the cell growth, the IC(50) value of bufalin for HL-60 cells was 0.025 micromol/L. (2) Apoptosis of HL-60 cells could be efficiently induced by bufalin at concentration of 0.010 micromol/L or higher. (3) Bufalin induced apoptosis of HL-60 cells in a dose- and time-dependent manner. (4) With G(1) phase cells decreasing, S phase cells increased, and then apoptotic cells increased with a diminution of S phase cells. (5) Bufalin-induced apoptosis of HL-60 cells was inhibited by ZnCl(2), an inhibitor of endonuclease, but not by cycloheximide, an inhibitor of protein synthesis. CONCLUSION: Bufalin could efficiently induce apoptosis of HL-60 cells, especially the S phase cells.